CN100575929C - Utilize one-dimensional microflow controlled biochip to detect the method for gene mutation in the cell - Google Patents
Utilize one-dimensional microflow controlled biochip to detect the method for gene mutation in the cell Download PDFInfo
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- CN100575929C CN100575929C CN200710034888A CN200710034888A CN100575929C CN 100575929 C CN100575929 C CN 100575929C CN 200710034888 A CN200710034888 A CN 200710034888A CN 200710034888 A CN200710034888 A CN 200710034888A CN 100575929 C CN100575929 C CN 100575929C
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Abstract
The invention discloses a kind of method of utilizing one-dimensional microflow controlled biochip to detect gene mutation in the cell, it is characterized in that making up sudden change detection chip based on the functionalization of one-dimensional microflow controlled micropearl array chip, total RNA or target DNA sample are diluted in the hybridization buffer that includes fluorescence probe, and under pressure-driven, flow through the microchannel and enter the functionalization micropearl array, wash-out is carried out in the high rigorous eluent injection channel that will contain formamide after the hybridization, observe and take bead surface fluorescence at last by the fluorescence inverted microscope, and with software the surface fluorescence intensity of particle is estimated, judge the mutation type of this detection site according to the difference of fluorescence intensity.The inventive method has solved the defective that characteristics such as trace detection, high sensitivity and high throughput analysis are difficult to and deposit in the gene mutation analysis method, and is expected to realize the gene mutation analysis on the unicellular level.
Description
Technical field
The present invention relates to the biological applications field of micro-total analysis system, relate in particular to the application and development of one-dimensional microflow controlled biochip in the cytogene mutation analysis.
Background technology
American and Britain, medium seven states have announced to finish the analysis first draft of human genomic sequence February calendar year 2001, make human biomedical research field change the genome times afterwards comprehensively over to from the genome epoch, i.e. functional genome's epoch.The geneticist generally believes the variation that the significant effort direction is analyst's genoid sequence of genome times afterwards comprehensively, because research human DNA sequence's sudden change, help the generation, development of more deep understanding human diseases and to the reaction of drug therapy, therefore the analyzing detecting method of sudden change just seems extremely important.Before 1985, utilize the Southern blotting, can filter out the mutant forms such as disappearance, insertion and frameshit reorganization of gene.For with the non-detectable sudden change of this method, can only use complicated time-consuming determined dna sequence analytic approach.Polymerase chain reaction (polymerase chain reaction, PCR) technology is the major progress in the mutation research, make the detection in Gene Mutation technology that significant progress arranged, but round pcr time-consuming, the false positive and the application of PCR method in sudden change detects that can not detect drawbacks limit such as a plurality of target sites simultaneously appear easily.The sudden change detection that appears as of micro-array chip (Microarray) provides a kind of ten minutes detection platform efficiently, directly realized the high flux and the information integration of mutation analysis, yet the analytical instrument of this method is very expensive, and detection sensitivity is low.Microflow control technique for the traceization that realizes sudden change and detect, rapid and high sensitivity provide may, but on the high flux performance, be still waiting to further develop.
Summary of the invention
The present invention aims to provide a kind of identification of cytogene sudden change and novel detection method of analysis of can be applicable to, solving the defective that the characteristics such as trace detection, high sensitivity and high throughput analysis that exist in the current gene mutation analysis method are difficult to and deposit, and promote further developing of this technical field.
The present invention is achieved through the following technical solutions the object of the invention:
Utilize one-dimensional microflow controlled biochip to detect the method for gene mutation in the cell, comprise that the silicon dioxide microballon activates with alkali on the one-dimensional microflow controlled biochip, carry out pre-service with biotin and Avidin again, the sudden change that adds biotin modification then respectively detects capture probe, with having modified the cell that silicon dioxide microballon that sudden change detects capture probe moves into described biochip microchannel one by one, form the functionalization micropearl array; Total RNA or target DNA sample are diluted in the hybridization buffer that includes fluorescence probe, and under pressure-driven, flow through the microchannel and enter the functionalization micropearl array, wash-out is carried out in the high rigorous eluent injection channel that will contain formamide after the hybridization, observe and take bead surface fluorescence at last by the fluorescence inverted microscope, and with software the surface fluorescence intensity of particle is estimated, judge the mutation type of this detection site according to the difference of fluorescence intensity.
The ratio of described total RNA and fluorescence probe is 1.5 * 10
7Gram: 1 mole, the ratio of DNA and fluorescence probe restrains more than or equal to 56: 1 mole; The flow velocity of hybridization buffer is 0.1 μ l/min under the described pressure-driven; The signal probe that described fluorescence probe is modified for the tetramethyl rhodamine; It is 60% formamide that the rigorous eluent of described height contains percent by volume.
Description of drawings
Fig. 1 is used to detect the process flow diagram of cytogene sudden change for one-dimensional microflow controlled biochip;
The sandwich hybridization illustraton of model of Fig. 2 for adopting among the present invention;
1-silicon dioxide microballon, the biotinylated bovine serum albumin(BSA) of 2-bag quilt,
The 3-streptavidin, the capture probe of 4-biotin modification,
The 5-sudden change detects the nucleic acid fragment of gene, the signal probe that 6-tetramethyl rhodamine is modified.
Fig. 3 detects the sensitivity result of gene mutation for one-dimensional microflow controlled biochip;
Fig. 4 detects the result of CNE2, A549 and three kinds of tumour cell p53 of SKBr-3 gene 17,5 bit codon mutation situations for one-dimensional microflow controlled biochip.
One-dimensional microflow controlled biochip adopts sandwich model as hybridization detection method, this modular concept is: as shown in Figure 2, biotinylated bovine serum albumin(BSA) 2 is attached on the solid phase interface silica microballon 1 by physisorption, strong combination occurs and is fixed on the silica bead surface with biotin in streptavidin 3, the capture probe 4 of biotin modification by the biotin modified and silica bead surface Avidin 3 specific bindings and be fixed on the silica microballon, complementary matching sequence in the specific selective recognition target nucleic acid sequence 5 of capture probe 4 energy of biotin modification, and the signal probe 6 that an other tetramethyl rhodamine is modified can mate combinations with target nucleic acid sequence 5, form the hybrid structure that is similar to sandwich after the triplicity, can realize the specific detection in the target nucleic acid sequence non-marked situation.
The advantage of the inventive method is: process of the present invention utilizes mRNA and DNA to be detected object, does not need the PCR process that adopts in the conventional method, thereby has avoided in the PCR process because pollute the false-positive situation that causes; Simultaneously because one-dimensional microflow controlled biochip has the double grading that trace detection and high flux detect, therefore the characteristics such as trace detection, high sensitivity and high throughput analysis are difficult to and the defective of depositing is solved preferably in the conventional mutation analysis method; The inventive method still can be distinguished the district of single base mutation under 40pM target level condition, detect when adopting single injected sampling just can carry out a plurality of target; The inventive method is in auxiliary lower hybridization and the elution process that can detect in real time and observe probe of CCD and computer software in addition, and be expected to use the inventive method to realize gene mutation analysis on the unicellular level, for generation, development and the personalized medicine of research human diseases provides a kind of efficient analytical method.
Embodiment
1. prepare sudden change detected silica microballon: the about 10～20mg of silicon dioxide microballon of cut-off footpath 40 μ m places the Ep pipe of 1.5ml, adds the NaOH activating surface 20min of 500 μ l 0.01M, cleans 3 times with TE, removes supernatant; The biotinylated bovine serum albumin(BSA) (Biotin-BSA) that adds 80 μ l, 0.1～1.0mg/ml places on the low temperature shaking table 4 ℃, and 350rpm sways 12～48h, cleans 3 times with TE then, and Biotin-BSA is attached to particle surface by physisorption; The streptavidin (Streptavidin) that adds 80 μ l, 0.1～1.0mg/ml places on the low temperature shaking table 4 ℃, and 350rpm sways 4～8h, and streptavidin can particle surface take place combine and be fixed on effectively with the biotin of particle surface, with TE cleaning 3 times; The sudden change that adds 50 μ l, 0.1～1.0 μ M biotin modification respectively detects capture probe Capture-C, Capture-G, Capture-T and Capture-A (as shown in table 1), and in 4 ℃, 350rpm sways 12～48h, and it is standby to clean back 4 ℃ of preservations with TE.Capture probe combines and is fixed on the particle with the Avidin specificity of particle surface by modified biotin, thereby has formed the functionalized SiO 2 particle with p53 detection in Gene Mutation ability.
2. prepare one-dimensional microflow controlled sudden change detection chip: dimethyl silicone polymer and hardening agent are fully mixed the back in vacuum pump, remove bubble, be tiled in then on the chip force plate, place 75 ℃ of baking oven 20～40min, waiting to solidify the back takes out, at last dimethyl silicone polymer (PDMS) sheet base is stripped down from force plate, include many cells that are used to hold microballon in this sheet base; This sheet base is placed on the inverted fluorescence microscope, by micrurgic method the functionalization microballon that being modified with sudden change detector probe of diameter about 40 μ m is placed in the cell in the sheet base microchannel: at first, use the burning pin of a cover import, draw pin and card grinding systems produce micromanipulation micro pipette, micro pipette is fixed on the micromanipulation system of inverted fluorescence microscope, then under micro-condition, different functionalization microballons is put into the cell of passage according to position encoded mode, and the corresponding different functionalization microballon of each cell is used for the multi-target detection of sample; Combine closely with clean slide and sheet base at last and finish the encapsulation of chip.
3. press the operating process of Fig. 1, to contain concentration and be 1pM～10nMp53-G sample and 10nM tetramethyl rhodamine and modify the hybridization buffer of signal probe (F-probe) (composition of described hybridization buffer is: 10mM kaliumphosphate buffer (pH7.6), 300mMNaCl, percent by volume are that 0.1% polysorbas20 and percent by volume are 30% formamide) flow velocity sudden change measuring ability micropearl array with 0.1 μ l/min under pressure-driven by placing in the one-dimensional biochip microchannel, behind the hybridization 20min, with the TE flushing channel of 5 μ l; Under pressure-driven, feed then and contain the high rigorous elution buffer that percent by volume is 60% formamide (composition of described elution buffer is: 10mM kaliumphosphate buffer (pH7.6), 150mMNaCl, percent by volume are that 0.1% polysorbas20 and percent by volume are 60% formamide) with the flow velocity of 2 μ l/min, utilize coupling (Capture-C and p53-G) and the single base (Capture-G that do not match, Capture-T, Capture-A and p53-G) the thermokinetics difference is carried out wash-out identification between sequence, uses the TE flushing channel behind the 5min once more.Reacted chip is placed under the CCD imaging system of fluorescence inverted microscope according to flow process shown in Figure 1, particle is observed shooting, and analyze, can obtain the result of sudden change detection sensitivity energy as shown in Figure 3 with the fluoroscopic image analysis software.The result shows, when target sequence concentration during at 0.04nM, signal to noise ratio (S/N ratio) is 4, and (signal to noise ratio (S/N ratio) is defined as the fluorescence signal that fluorescence signal that complete matching hybridization produced produces divided by single base mismatch hybridization, think that simultaneously the fluorescence signal that has only complete matching hybridization to be produced is effective greater than 4 o'clock data divided by the fluorescence signal that single base mismatch hybridization produces), raising along with target sequence concentration, the fluorescence of matching hybridization generation also improves constantly fully, when target sequence concentration reaches 3nM, fully the fluorescence signal that produces of matching hybridization reaches maximum, this shows utilize the inventive method can be under 0.04nM target sequence concentration the identification form base mutation.
The oligonucleotide probe that is used to detect the p53 gene mutation that table 1 is synthetic
| Code | Describe | Sequence |
| ?Capture-C | Identification point is the capture probe of C | ?5’-GGGCAG CGCCTCACAACCAAAAAAAAAA-Biotin-3’ |
| ?Capture-G | Identification point is the capture probe of G | ?5’-GGGCAG GGCCTCACAACCAAAAAAAAAA-Biotin-3’ |
| ?Capture-T | Identification point is the capture probe of T | ?5’-GGGCAG TGCCTCACAACCAAAAAAAAAA-Biotin-3’ |
| ?Capture-A | Identification point is the capture probe of A | ?5’-GGGCAG AGCCTCACAACCAAAAAAAAAA-Biotin-3’ |
| ?p53-G | P53 gene target sequence | ?5’-GGTTGTGAGGC GCTGCCCAAGCGAGCACTG ?CCCAACAACA?CCAGC-3’ |
| ?F-probe | Signal probe | ?5’TAMRA-AAAAAAAAAAGCTGGTGTTGTTGGGCAGTG ?CTCGCTT-3’ |
1. prepare sudden change detected silica microballon: the about 10～20mg of silicon dioxide microballon of cut-off footpath 40 μ m places the Ep pipe of 1.5ml, adds the NaOH activating surface 20min of 500 μ l 0.01M, cleans 3 times with TE, removes supernatant; The biotinylated bovine serum albumin(BSA) (Biotin-BSA) that adds 80 μ l, 0.1～1.0mg/ml places on the low temperature shaking table 4 ℃, and 350rpm sways 12～48h, cleans 3 times with TE then, and Biotin-BSA is attached to particle surface by physisorption; The streptavidin (Streptavidin) that adds 80 μ l, 0.1～1.0mg/ml places on the low temperature shaking table 4 ℃, and 350rpm sways 4～8h, and streptavidin can particle surface take place combine and be fixed on effectively with the biotin of particle surface, with TE cleaning 3 times; The sudden change that adds the biotin modification of 50 μ l, 0.1～1.0 μ M respectively detects capture probe Capture-C, Capture-G, Capture-T and Capture-A (as shown in table 1), and in 4 ℃, 350rpm sways 12～48h, and it is standby to clean back 4 ℃ of preservations with TE.Capture probe combines and is fixed on the particle with the Avidin specificity of particle surface by modified biotin, thereby has formed the functionalized SiO 2 particle with p53 detection in Gene Mutation ability.
2. prepare one-dimensional microflow controlled sudden change detection chip: dimethyl silicone polymer and hardening agent are fully mixed the back in vacuum pump, remove bubble, be tiled in then on the chip force plate, place 75 ℃ of baking oven 20～40min, waiting to solidify the back takes out, at last PDMS sheet base is stripped down from force plate, include many cells that are used to hold microballon in this sheet base.To be placed on the inverted fluorescence microscope with the one-dimensional microflow controlled biochip sheet base of dimethyl silicone polymer casting, by micrurgic method the functionalization microballon that being modified with sudden change detector probe of diameter about 40 μ m is placed in the cell in the microchannel: at first, use the burning pin of a cover import, draw pin and card grinding systems produce micromanipulation micro pipette, micro pipette is fixed on the micromanipulation system of inverted fluorescence microscope, then under micro-condition, different functionalization microballons is put into the cell of passage according to position encoded mode, and the corresponding different functionalization microballon of each cell is used for the multi-target detection of sample; Combine closely with clean slide and sheet base at last and finish the encapsulation of chip.
3. carry out quantitatively to these three kinds of cells difference extracted total RNA of nasopharyngeal carcinoma cell (CNE2), lung carcinoma cell (A549) and breast cancer cell (SKBr-3), and by the uv absorption method; Getting the total RNA of 3 μ g (described three kinds of cells) (described hybridization buffer composition is: 10mM kaliumphosphate buffer (pH7.6), 300mMNaCl, percent by volume are that 0.1% polysorbas20 and percent by volume are 30% formamide) in the hybridization buffer of 20 μ l respectively dilutes, adding the signal probe of tetramethyl rhodamine modification simultaneously and making its final concentration in hybridization buffer is 10nM; Then under pressure-driven with the flow velocity of 0.1 μ l/min by the functionalization micropearl array in the one-dimensional biochip, behind the hybridization reaction 20min, with the TE flushing channel and under pressure-driven with high rigorous elution buffer wash-out 5min (composition of described elution buffer is: 10mM kaliumphosphate buffer (pH7.6), 150mMNaCl, percent by volume are that 0.1% polysorbas20 and percent by volume are 60% formamide), flow velocity is 2 μ l/min; Reacted chip is placed under the CCD imaging system of fluorescence inverted microscope, particle is observed shooting, and analyze, can obtain the result of sudden change detection sensitivity energy as shown in Figure 4 with the fluoroscopic image analysis software.The result shows, has only the SKBr-3 cell that (the base conversion of CGC＞CAC), and 175 bit codons of other two kinds of cell p53 genes are not undergone mutation, and still are wild type has taken place in three kinds of tumour cells.This result is consistent with traditional sequencing result.
Claims (3)
1. method of utilizing one-dimensional microflow controlled biochip to detect gene mutation in the cell, comprise that the silicon dioxide microballon activates with alkali on the one-dimensional microflow controlled biochip, carry out pre-service with biotin and Avidin again, the sudden change that adds biotin modification then respectively detects capture probe, the silicon dioxide microballon of having modified sudden change detection capture probe is moved into one by one the cell of described biochip microchannel, form the functionalization micropearl array, it is characterized in that total RNA or target DNA sample are diluted in the hybridization buffer that includes fluorescence probe, and under pressure-driven, flow through the microchannel and enter the functionalization micropearl array with the flow velocity of 0.1 μ l/min, wash-out is carried out in the high rigorous eluent injection channel that will contain formamide after the hybridization, the volume fraction of described formamide in the rigorous eluent of height is 60%, observe and take bead surface fluorescence at last by the fluorescence inverted microscope, and with software the surface fluorescence intensity of particle is estimated, judge the mutation type of this detection site according to the difference of fluorescence intensity.
2. the method for utilizing one-dimensional microflow controlled biochip to detect gene mutation in the cell according to claim 1, the ratio that it is characterized in that described total RNA and fluorescence probe is 1.5 * 10
7Gram: 1 mole, the ratio of DNA and fluorescence probe restrains more than or equal to 56: 1 mole.
3. the method for utilizing one-dimensional microflow controlled biochip to detect gene mutation in the cell according to claim 1 and 2 is characterized in that the signal probe that described fluorescence probe is modified for the tetramethyl rhodamine.
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|---|---|---|---|---|
| CN101520417B (en) * | 2009-04-02 | 2012-02-15 | 深圳市杰普特电子技术有限公司 | Fluorimetric biochip |
| CN101709261B (en) * | 2009-12-11 | 2013-06-19 | 香港城市大学深圳研究院 | Microfluidic microbead array chip and application thereof in virus analysis |
| WO2011150675A1 (en) * | 2010-06-01 | 2011-12-08 | 厦门大学 | Biochip comprising multiple microchannels |
| CN103013821B (en) * | 2012-12-03 | 2014-11-26 | 清华大学 | Avidin-biotin system cell patterning-based chip and preparation and applications thereof |
| CN103333967B (en) * | 2013-07-12 | 2016-06-22 | 湖南工程学院 | A kind of nucleic acid detection method based on microfluidic microbead array chip |
| CN103529195B (en) * | 2013-10-24 | 2014-08-06 | 山东大学 | Detection method applied to measurement of trace target materials |
| CN104894264A (en) * | 2015-06-04 | 2015-09-09 | 中国科学院海洋研究所 | New method for detecting sulfate reducing bacteria |
| US20170205404A1 (en) * | 2016-01-19 | 2017-07-20 | General Electric Company | Multifunctional beads and methods of use for capturing rare cells |
| CN111735801B (en) * | 2019-03-25 | 2021-09-28 | 南京大学 | Fluorescence analysis method based on HCR and cation exchange reaction of hydrogel |
-
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Non-Patent Citations (8)
| Title |
|---|
| A novel sandwich assay with molecular beacon as reportprobe for nucleic acids detection on one-dimensionalmicrofluidic beads array. Xinbing Zuo等.Analytica Chimica Acta,Vol.587 . 2007 |
| A novel sandwich assay with molecular beacon as reportprobe for nucleic acids detection on one-dimensionalmicrofluidic beads array. Xinbing Zuo等.Analytica Chimica Acta,Vol.587 . 2007 * |
| 一维微流控微珠阵列芯片用于肿瘤转移相关基因表达谱检测. 文建辉等.科学通报，第52卷第6期. 2007 |
| 一维微流控微珠阵列芯片用于肿瘤转移相关基因表达谱检测. 文建辉等.科学通报，第52卷第6期. 2007 * |
| 微流控芯片实验室在基因分析研究中的应用. 秦建华等.色谱，第21卷第5期. 2003 |
| 微流控芯片实验室在基因分析研究中的应用. 秦建华等.色谱，第21卷第5期. 2003 * |
| 线性微珠阵列生物芯片用于P53蛋白功能相关高突变位点的检测. 张何等.第四届全国化学生物学学术会议暨国际化学与生物/医学交叉研讨会. 2005 |
| 线性微珠阵列生物芯片用于P53蛋白功能相关高突变位点的检测. 张何等.第四届全国化学生物学学术会议暨国际化学与生物/医学交叉研讨会. 2005 * |
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